Bifunctional Magnetic Supramolecular-Organic Framework: A Nanoprobe for Simultaneous Enrichment of Glycosylated and Phosphorylated Peptides

Abstract

Protein glycosylation and phosphorylation are two important protein post-translational modifications. Mass spectrometry (MS) has been proved to be a powerful technique in comprehensive characterization of protein glycosylation and phosphorylation; however, the complexity of biological matrices and weak ionization efficiency bring a big challenge. Capturing glycopeptides and phosphopeptides from complicated biological samples is indispensable before MS determinations. In this study, a bifunctional gallium ion immobilized magnetic pertriflated pillar[5]arene supramolecular-organic framework (magOTfP5SOF-Ga 3+ ) was designed for the one-step simultaneous enrichment of glycopeptides and phosphopeptides. Thanks to the abundant sulfonic acid groups, the material owns strong hydrophilicity and leads to hydrophilic interaction chromatography for glycopeptides enrichment. Simultaneously, the high loading amount of gallium ion provides immobilized metal ion affinity for phosphopeptides enrichment. The established platform possesses quick magnetic response performance, high sensitivity (detection limits as low as 0.1 fmol and 0.05 fmol for glycopeptides and phosphopeptides, respectively), and good reusability. In addition, the method was applied to the determination of glycopeptides and phosphopeptides in clinical specimens, cell lysates, and mouse liver tissue samples, demonstrating its highly sensitive and specific glycoproteomics and phosphorproteomics analysis in complex biosamples.

Facile Preparation of Chitosan‐modified Mesoporous Titanium Dioxide Film on Fused‐silica Capillary for Selective Enrichment of Phosphopeptides

“…The enrichment performance of the Cts/mTiO 2 film is better than the previously reported enrichment materials (Table 1). [59–65] These results fully validate Cts/mTiO 2 film can serve as a reliable and efficient material to enrich and separate phosphopeptides.…”

Preparation of IgY Oriented Conjugated Fe3O4 MNPs as Immunomagnetic Nanoprobe for Increasing Enrichment Efficiency of Staphylococcus aureus Based on Adjusting the pH of the Solution System

“…The Fourier-transform infrared (FT-IR) spectra of Fe 3 O 4 MNPs and nanoprobe are shown in Figure 2D . In the spectrum of bare Fe 3 O 4 MNPs, the characteristic band at 587, 1,050, 1,384, 1,640, and 3,310 cm −1 could be attributed to the stretching vibration of the Fe-O bond, C-N bond, C-H bond, C-O bond, C=O bond, and O-H bond, respectively ( 11 , 36 ). After conjugation, the new peaks at 1,536 cm −1 were attributed to the N-H stretching vibration ( 37 ), indicating the occurrence of amino-carboxyl reactions on the surface of MNPs and the formation of peptide bonds.…”

A Pragmatic Guide to Enrichment Strategies for Mass Spectrometry–Based Glycoproteomics

“…Of importance, others have shown that sialylated N-glycopeptides can be enriched not only by TiOx but also by Fe 3+ -IMAC ( 176 ) and Ti 4+ -IMAC ( 177 ). Coenrichment of phosphopeptides and glycopeptides can also be achieved with hydrophilic interaction chromatography (HILIC, see below) ( 178 , 179 ) and nanomaterials functionalized with HILIC matrices, metal ions like Ti 4+ or Ga 3+ , or both ( 180 , 181 , 182 ).…”

“…A common property achieved by these materials is hydrophilicity, although the avenues to achieve such properties differ. Materials include cobalt sulfide nanoleaves ( 242 ), covalent/metal organic framework approaches ( 181 , 243 , 244 , 245 , 246 ), carbon microspheres ( 247 ), and polymeric monoliths functionalized with piperazine ( 248 ), among others. As described above, some of these materials can have bifunctional properties to enable HILIC-based glycopeptide enrichment and metal ion–based phosphopeptide enrichment ( 181 , 182 , 249 ).…”